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Maruthamuthu, Murali kannan,Selvamani, Vidhya,Nadarajan, Saravanan Prabhu,Yun, Hyungdon,Oh, You-Kwan,Eom, Gyeong Tae,Hong, Soon Ho Springer-Verlag 2018 Journal of industrial microbiology & biotechnology Vol.45 No.1
<P>In a cell-surface display (CSD) system, successful display of a protein or peptide is highly dependent on the anchoring motif and the position of the display in that anchoring motif. In this study, a recombinant bacterial CSD system for manganese (Mn) and cobalt (Co) recovery was developed by employing OmpC as an anchoring motif on three different external loops. A portion of Cap43 protein (TRSRSHTSEG)(3) was employed as a manganese and cobalt binding peptide (MCBP), which was fused with OmpC at three different external loops. The fusions were made at the loop 2 [fusion protein-2 (FP2)], loop 6 (FP6), and loop 8 (FP8) of OmpC, respectively. The efficacy of the three recombinant strains in the recovery of Mn and Co was evaluated by varying the concentration of the respective metal. Molecular modeling studies showed that the short trimeric repeats of peptide probably form a secondary structure with OmpC, thereby giving rise to a difference in metal recovery among the three recombinant strains. Among the three recombinant strains, FP6 showed increased metal recovery with both Mn and Co, at 1235.14 (1 mM) and 379.68 (0.2 mM) mu mol/g dry cell weight (DCW), respectively.</P>
Maruthamuthu, Murali kannan,Hong, Jiyeon,Arulsamy, Kulandaisamy,Somasundaram, Sivachandiran,Hong, SoonHo,Choe, Woo-Seok,Yoo, Ik-Keun Springer-Verlag 2018 Bioprocess and biosystems engineering Vol.41 No.4
<P>Peptide-displaying Escherichia coli cells were investigated for use in adsorptive removal of bisphenol A (BPA) both in Luria-Bertani medium including BPA or ATM thermal paper eluted wastewater. Two recombinant strains were constructed with monomeric and dimeric repeats of the 7-mer BPA-binding peptide (KSLENSY), respectively. Greater than threefold increased adsorption of BPA [230.4 A mu mol BPA per g dry cell weight (DCW)] was found in dimeric peptide-displaying cells compared to monomeric strains (63.4 A mu mol per g DCW) in 15 ppm BPA solution. The selective removal of BPA from a mixture of BPA analogs (bisphenol F and bisphenol S) was verified in both monomeric and dimeric peptide-displaying cells. The binding chemistry of BPA with the peptide was assumed, based on molecular docking analysis, to be the interaction of BPA with serine and asparagine residues within the 7-mer peptide sequence. The peptide-displaying cells also functioned efficiently in thermal paper eluted wastewater containing 14.5 ppm BPA.</P>
홍순호,( Murali Kannan Maruthamuthu ),석정욱,정재훈 한국공업화학회 2016 한국공업화학회 연구논문 초록집 Vol.2016 No.1
Chimeric sensor kinase (SK)-based biosensor was constructed using Pseudomonas putida AauS. The AauZ chimera SK was constructed by integration of the sensing domain of AauS with the catalytic domain of EnvZ to control the expression of the ompC gene in response to acidic amino acids. Real-time quantitative PCR and GFP fluorescence studies showed increased ompC gene expression and GFP fluorescence as the concentration of acidic amino acids increased. These data suggest that AauS-based recombinant E. coli can be used as a bacterial biosensor of acidic amino acids. By employing the chimeric SK strategy, various bacteria biosensors for use in the development of biochemical-producing recombinant microorganisms can be constructed. <sup>**</sup>This work was supported by a grant from the Next-Generation Bio- Green 21 Program (SSAC, grant number: PJ011116) by RDA and Basic Science Research Program by the Ministry of Education (NRF-2014R 1A1A2054726).
Irisappan Ganesh,Murali Kannan Maruthamuthu,유익근,홍순호 한국생물공학회 2015 Biotechnology and Bioprocess Engineering Vol.20 No.5
A positive feedback loop was introduced to modify the dynamic behavior of fumarate sensing DcuSZ chimera TCS. To construct the positive feedback loop, the ompR gene was cloned downstream of the ompC promoter. The ompC promoter induced the expression of OmpR, which in turn induced the expression of the ompC promoter. Through the introduction of this positive feedback loop, the transcriptional expression levels of ompC increased 2.6-fold. When GFP was used as a reporter protein, a 64% increase in fluorescence level was observed. These results suggest that sensitivity of the TCS based fumarate sensing system can be engineered through the introduction of a positive feedback loop.
Vidhya Selvamani,Murali Kannan Maruthamuthu,Kulandaisamy Arulsamy,엄경태,홍순호 한국화학공학회 2017 Korean Journal of Chemical Engineering Vol.34 No.6
Methylobacterium organophilum XX is a type II facultative methylotroph that can grow on methanol. In M. organophilum XX, the MxcQ/MxcE two-component system (TCS) is involved in methanol metabolism. EnvZ/OmpR in E. coli TCS was exploited to develop a methanol biosensor by engaging the MxcQ/MxcE TCS system. The MxcQZ/ OmpR methanol sensing chimeric TCS was constructed by integrating the sensing domain of M. organophilum MxcQ with the transmitter domain of E. coli EnvZ. The response regulator of the chimeric TCS system is OmpR, which regulates the expression of the ompC and gfp. The expression of ompC was monitored by real-time quantitative PCR analysis. The expression of gfp also confirmed the expression of the ompC. The maximum expression of ompC and gfp occurred with 0.05% of methanol, and the expression started to decline with further increases in methanol concentration. This system delivers rapid detection of methanol in the environment.
( Sivachandiran Somasundaram ),( Murali Kannan Maruthamuthu ),( Irisappan Ganesh ),( Gyeong Tae Eom ),( Soon Ho Hong ) 한국미생물생명공학회(구 한국산업미생물학회) 2017 Journal of microbiology and biotechnology Vol.27 No.9
Gamma-aminobutyric acid is a precursor of nylon-4, which is a promising heat-resistant biopolymer. GABA can be produced from the decarboxylation of glutamate by glutamate decarboxylase. In this study, a synthetic scaffold complex strategy was employed involving the Neurospora crassa glutamate decarboxylase (GadB) and Escherichia coli GABA antiporter (GadC) to improve GABA production. To construct the complex, the SH3 domain was attached to the N. crassa GadB, and the SH3 ligand was attached to the N-terminus, middle, and C-terminus of E. coli GadC. In the C-terminus model, 5.8 g/l of GABA concentration was obtained from 10 g/l glutamate. When a competing pathway engineered strain was used, the final GABA concentration was further increased to 5.94 g/l, which corresponds to 97.5% of GABA yield. With the introduction of the scaffold complex, the GABA productivity increased by 2.9 folds during the initial culture period.
SAMBANDAM RAVIKUMAR,Irisappan Ganesh,Murali Kannan Maruthamuthu,홍순호 한국화학공학회 2015 Korean Journal of Chemical Engineering Vol.32 No.10
In an attempt to create an acidic amino acid-sensing Escherichia coli, a chimeric sensor kinase (SK)-based biosensor was constructed using Pseudomonas putida AauS. AauS is a sensor kinase that ultimately controls expression of the aau gene through its cognate response regulator AauR, and is found only in P. putida KT2440. The AauZ chimera SK was constructed by integration of the sensing domain of AauS with the catalytic domain of EnvZ to control the expression of the ompC gene in response to acidic amino acids. Real-time quantitative PCR and GFP fluorescence studies showed increased ompC gene expression and GFP fluorescence as the concentration of acidic amino acids increased. These data suggest that AauS-based recombinant E. coli can be used as a bacterial biosensor of acidic amino acids. By employing the chimeric SK strategy, various bacteria biosensors for use in the development of biochemicalproducing recombinant microorganisms can be constructed.